Application Details

Application Details

ShowHide

Application Notes

Western Blot 1:2000-1:10000, Immunohistochemistry 1:10-1:500, Immunoprecipitation 2-5 μL, Immunohistochemistry-Paraffin 1:10-1:500This VEGF Receptor 2 antibody is useful for Immunohistochemistry paraffin-embedded sections (PMID 10751359), Immunoprecipitation and Western blot where a doublet can been seen at ~ 150 kDa. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol

Protocol specific for VEGF Receptor 2 Antibody Western Blot Protocol1. Perform SDS-PAGE (3-8 %) on samples to be analyzed, loading 20 µg of total transfected protein per lane.. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.. Stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.. Rinse the blot in TBS for approximately 5 minutes.. Block the membrane using 5 % non-fat dry milk in TBS with 0.5 % BSA for 1 hour.. Dilute the rabbit anti-VEGFR2 primary antibody in blocking buffer and incubate 1.5 hours at room temperature.. Wash the membrane in water for 5 minutes and apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.. Wash the blot in TBS containing 0.05-0.1 % Tween-20 for 10-20 minutes.. Wash the blot in type I water for an additional 10-20 minutes (this step can be repeated as required to reduce background).. Apply the detection reagent of choice in accordance with the manufacturer's instructions (Amersham's ECL is the standard reagent used).Note: Tween-20 can be added to the blocking buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding.